生物学杂志

生物学杂志 ›› 2023, Vol. 40 ›› Issue (4): 31-.doi: 10.3969/j.issn.2095-1736.2023.04.031

• 研究报告 • 上一篇    下一篇

FoxM1靶向肽9R-P49对L929成纤维细胞的抑制作用及机制

常　苗， 项　坤， 何佳萌， 梁安平， 花欣怡， 刘新荣， 江育虹， 茆灿泉   

  1. 西南交通大学 生命科学与工程学院， 成都 610031
  • 出版日期:2023-08-18 发布日期:2023-08-10
  • 通讯作者: 茆灿泉，博士，教授，研究方向为生物医药研发，E-mail:maocq@swjtu.edu.cn
  • 作者简介:常苗，硕士研究生，研究方向为肿瘤靶向多肽药物，E-mail:1624953557@qq.com
  • 基金资助:
    国家自然科学基金资助项目（No.81872789）；成都市重点研发支撑计划项目（No.2018-YF05-00004-SN）

Inhibition and preliminary mechanisms of a FoxM1-targeting polypeptide 9R-P49 on fibroblast L929 cells

CHANG Miao, XIANG Kun, HE Jiameng, LIANG Anping, HUA Xinyi, LIU Xinrong,JIANG Yuhong, MAO Canquan   

  1. School of Life Science and Engineering, Southwest Jiaotong University, Chengdu 610031, China
  • Online:2023-08-18 Published:2023-08-10

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摘要： 为探究9R-P49多肽对FoxM1的作用及其对成纤维细胞的调控机制，采用CCK-8检测细胞抑制率、AO/EB双染和流式细胞术检测细胞凋亡、Transwell检测细胞迁移、细胞平板克隆检测细胞增殖；使用PyMOL分子对接软件预测P49多肽与FoxM1-DBD的潜在结合位点；采用0、30.0和60.0 μg/mL的9R-P49多肽处理小鼠成纤维L929细胞，Western Blot检测FoxM1蛋白的表达，最后通过RNA转录组分析差异基因及相关通路。结果表明：在L929细胞中9R-P49下调FoxM1蛋白的表达，同时，9R-P49能够抑制细胞增殖和迁移、促进细胞凋亡；P49多肽与FoxM1-DBD上的Arg 297、Ser 290和Asp 293位点存在潜在结合；给药处理后转录组差异基因GO分类主要富集于细胞过程、单一组织过程、生物调节过程和新陈代谢过程等，KEGG分类主要富集于免疫系统、内分泌系统、信号转导和新陈代谢等通路。在成纤维L929细胞中9R-P49能够通过调控FoxM1表达抑制L929细胞增殖和迁移并促进凋亡。
关键词　9R-P49； FoxM1； 纤维化； 转录组； 多肽

关键词: 9R-P49, FoxM1, 纤维化, 转录组, 多肽

Abstract: To explore the regulatory effect of 9R-P49 peptide on FoxM1 and its preliminary mechanism on fibroblast L929 cells, CCK-8 was used to detect cell inhibition rate, AO/EB double staining and flow cytometry to detect cell apoptosis, transwell assay to detect cell migration and cell plate clone to detect cell proliferation. The potential binding sites of the P49 peptide to FoxM1-DNA binding domain（FoxM1-DBD） were predicted using PyMOL molecular docking software. Mouse fibroblast L929 cells were treated with 0, 30.0 and 60.0 μg/mL 9R-P49 peptide and the expression of FoxM1 protein was detected by Western Blot. Finally, the differential expressed genes and related pathways were analyzed by RNA transcriptome. The results showed that 9R-P49 could inhibit FoxM1 protein expression in L929 cells. At the same time, 9R-P49 could inhibit cell proliferation and promote cell apoptosis of L929 cells. P49 peptide binds potentially to the Arg 297, Ser 290 and Asp 293 sites on FoxM1-DBD. The GO classification of differentially expressed genes in the transcriptome after drug treatment was mainly enriched in cellular process, single-organism process, biological regulation, and metabolic process. The KEGG classification was mainly enriched in the immune system, endocrine system, signal transduction, and metabolism pathways. In conclusion, 9R-P49 can inhibit the proliferation of L929 cells and promote apoptosis by regulating the expression of FoxM1 in L929 cells.

Key words: 9R-P49, FoxM1, fibrosis, transcriptome, polypeptide

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